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Western blot analysis of Morgue-associated proteins confirms Morgue association with polyubiquitin and SkpA.

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posted on 2013-09-30, 01:56 authored by Ying Zhou, Yiqin Wang, Barbara A. Schreader, John R. Nambu

A. Silver stained SDS gel of proteins from P[da-Gal4], P[UAS-Morgue] or P[da-Gal4],P[UAS-Morgue-3xFLAG] adult flies. Note that apparent molecular weight of Morgue-3xFLAG (arrowhead) is greater than native Morgue. Note smaller proteins (lower arrowheads) that are present in Morgue-3xFLAG expressing flies. B. Anti-SkpA Western blot of proteins purified from P[da-Gal4], P[UAS-Morgue] or P[da-Gal4], P[UAS-Morgue:3xFlag] flies via anti-FLAG chromatography. Note prominent SkpA band (arrowhead) present in proteins purified from P[da-Gal4], P[UAS-Morgue:3xFlag]. A shared background band of approximately 30 kDa is present in both samples. C. Anti-ubiquitin Western blot of proteins purified from P[da-Gal4], P[UAS-Morgue] or P[da-Gal4], P[UAS-Morgue:3xFlag] flies via anti-FLAG chromatography. Note ubiquitin band (arrowhead) present in proteins purified from P[da-Gal4], P[UAS-Morgue:3xFlag].

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